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FEATURES

FEATURES
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2448

Microbial biotechnology
Arnold L. Demain
For thousands of years, microorganisms have been used to supply products such as bread, beer and wine. A second phase
of traditional microbial biotechnology began during World War I and resulted in the development of the acetone-butanol and
glycerol fermentations, followed by processes yielding, for example, citric acid, vitamins and antibiotics. In the early
1970s, traditional industrial microbiology was merged with molecular biology to yield more than 40 biopharmaceutical
products, such as erythropoietin, human growth hormone and interferons. Today, microbiology is a major participant in global
industry, especially in the pharmaceutical, food and chemical industries.

icroorganisms are important for many
reasons, particularly because they produce
things that are of value to us1. These can be
very large materials
(e.g. proteins, nucleic acids, carbohydrate
polymers, even cells) or smaller molecules and are
usually divided into metabolites that are essential for
vegetative growth (primary) and those that are
inessential (secondary).
Although microbes are extremely good at
producing an amazing array of valuable products,
they usually produce these compounds in small
amounts that are needed for their own benefit.
Regulatory mechanisms have evolved that enable a
strain to avoid excessive pro- duction of its
metabolites so that it can compete effi- ciently with
other forms of life and survive in nature. By
contrast, the industrial microbiologist screens for a
‘wasteful’ strain that will overproduce a particular
com- pound that can be isolated and marketed.
After a desired strain has been found, a development
program is initiated to improve titers by modification
of culture conditions using mutation
and
recombinant DNA techniques. The main reason for
the use of microor- ganisms to produce compounds
that can otherwise be isolated from plants and
animals, or synthesized by chemists, is the ease of
increasing production by envi- ronmental and genetic
manipulation; 1000-fold increases have been recorded
for small metabolites2.

M

Traditional microbial biotechnology
Primary metabolites
Primary metabolites are the small molecules of living cells; they are intermediates or end products of
the
pathways of intermediary metabolism, building
blocks
for essential macromolecules, or are converted into

A.L. Demain (demain@mit.edu) is at the
Biology Department, Massachusetts Institute of
Technology, Cambridge, MA 02139, USA.

lactic). gellan).2 billion pounds of monosodium glutamate are made annually by fermentation using various species of the genera Corynebacterium (e. B. polyols (glycerol. biotin]. sorbose) and vitamins [riboflavin (B2). threonine. the major commercial amino acid. glutamicum) and Brevibacterium (e. Another method is to produce mutants that are resistant to a toxic analog of the desired metabolite. an antimetabolite.cinic. which is very important in the production of 26 FEATURES acid. However. Molar yields of glutamate from sugar are 50–60% and broth concentrations reach over 100 g l21. All rights reserved. fumaric. flavum and B.FEATURESPrimary metabolites used in the food and feed coenzymes. feedback regulation is bypassed by isolating auxotrophic mutants and partially starving them of their requirements. sugars (fructose. Mutants During amino acid production. ribose. thus allowing its biosynthesis 0167-7799/00/$ – see front matter © 2000 Elsevier Science Ltd. suc. tryptophan). organic acids (acetic. phenylala. xylitol). cyanocobalamin (B12). C. 18) . lysine.g. glutamic acid overproduction would not occur because of feedback regulation. 59-inosinic acid). PII: S0167-7799(99)01400-6 TIBTECH JANUARY 2000 (Vol. lactofermentum).nine. modification of the cell membrane can cause glutamate to be pumped out of the cell. erythritol. Approximately 1. propionic.g. L-glutamic Glutamic acid Normally. amino acids (monosodium glutamate. Fermentation Another factor is the increase in outward permeability. that is. polysaccharides (xanthan. Combinations of auxotrophic and antimetabolite resistance mutations are common in primary metaboliteproducing microorganisms. flavor nucleotides (59-guanylic acid. industries include: alcohols (ethanol). mannitol.

US$450 million for L-lysine. all of these manipulations result in a phospholipid. This pathway is controlled very tightly in an organism such as Escherichia coli. In addition to the difference in the mode of aspartate kinase feedback inhibition. a glutamate-producing wild-type Corynebacterium is converted into a lysine-overproducing mutant that cannot grow unless methionine and threonine are added to the medium. Excretion uses a (2OH2)–lysine symporter and is catalysed by a dipeptide-uptake system dependent on electromotive force. have been major influences in constructing bacterial strains capable of producing these levels (in g l21) . which includes three aspartate kinases that are each regulated by a different end product. Recombinant technology and traditional mutagenesis.g. oleate limitation of oleate auxotrophs. US$198 million for L-phenylalanine and US$43 million for L-aspartate. As a result. By the genetic removal of homoserine dehydrogenase. In addition. Apparently. Lysin e Most cereals are deficient in the essential amino acid L-lysine.35 g L-lysine • HCl g21 glucose used (molar yield of 0. However. the lysine gradient and the proton gradient. not on ATP. lysine overproducers differ from E. The excretion is carried out by a specific efflux system involving a carrier that is dependent on membrane potential. The major manipulation for lysine production is aimed at increasing the levels of feedback-resistant aspartate kinase and dihydrodipicolinate synthase.deficient cytoplasmic membrane. (2) the first and second enzymes of the lysine branch (dihydrodipicolinate synthetase and dihydrodipicolinate reductase) are neither inhibited nor repressed by lysine in lysine overproducers. lysine industrial production yields 120 g l21 and 0. World markets for amino acids amount to US$915 million for L-glutamate. glutamicum and its relatives). in lysine-fermentation organisms (e.25–0. Lysine excretion is via active transport involving a carrier and is driven by membrane potential. plus selection. As long as the threonine supplement is maintained at a limiting concentration. glycerol limitation of glycerol auxotrophs. the initial enzymes are inhibited by their respective end products and no overproduction occurs.35 mol of L-lysine mol21 of glucose used). various mutants of C. or addition of penicillin or fatty acid derivatives to exponentially growing cells. the intracellular concentration of threonine is the limiting factor and feedback inhibition of aspartate kinase is bypassed. after each branch point.to proceed unabated. Lysine is a member of the aspartate family of amino acids and is produced in bacteria by a branched pathway that also produces methionine. threonine and isoleucine. coli in the following ways: (1) no feedback repression of aspartate kinase or aspartate semialdehyde dehydrogenase occurs in lysine overproducers. there is only a single aspartate kinase. and (3) L-lysine decarboxylase is absent in lysine overproducers. This membrane alteration is intentionally effected by biotin limitation (all glutamic acid bacteria are biotin auxotrophs). Recombinant DNA technology Recombinant DNA technology is beginning to have a major impact on amino acid production3.4.25–0. which is regulated via concerted feedback inhibition by threonine and lysine.

A high level of citric acid production is also associated with a high intracellular concentration of fructose 2. with biotin 5-adenylate acting as corepressor. L-arginine.0) lead to the production of 27 . Approximately 2500 tons of GMP and IMP are produced annually in Japan alone. Fungi Filamentous fungi are widely used for the commercial production of organic acids.ronment that represses the formation of the early enzymes in the pathway. namely guanylic acid (GMP) and inosinic acid (IMP). feedback repression is caused by the enzyme protein acetyl-coenzyme A car. an activator of glycolysis. Eremothecium ashbyii and Ashbya gossypii. Lproline. 28. cobinamide. 3.6biphosphate. for example. the entire process is carried out under low oxygen. Vitamin B12 is produced on an industrial scale by Propionibacterium shermanii or Pseudomonas denitrificans. During production of biotin. and molecular cloning can produce 600 mg l21 in the presence of high con.centrations of sulfur and ferrous iron. Lisoleucine. selection for resist. which synthesize riboflavin in concentrations greater than 20 g l21. The early stage of the P. with a world market of US$350 million.6-dimethylbenzimidazole.g.of amino acids: L-threonine. 31. 1 billion pounds of citric acid are produced per year with a market value of US$1.manii fermentation is conducted under anaerobic con. Such a titer is high enough to economically compete with the traditional chemical production process.boxylase biotin holoenzyme synthetase. 55. and Lhistidine.ance to biotin antimetabolites. A high level of oxygen results in an oxidizing intracellular envi. Strains of Serratia marcescens obtained by mutagenesis. Commercial interest in nucleotide fermentations is due to the activity of two purine ribonucleoside 59-monophosphates. as flavor enhancers.0). 100. 100. sher. Production of vitamin B12 has reached levels of 150 mg l21 and a world market value of US$71 million. The commercial process uses Aspergillus niger in media deficient in iron and manganese. 26.7–2. Other factors contributing to high citric acid production are the inhibition of isocitrate dehydrogenase by citric acid and the low optimum pH (1. New processes using Candida sp. The culture is TIBTECH JANUARY 2000 (VOL 18) then aerated and dimethylbenzimidazole is added.ditions in the absence of the precursor 5. 34. In the P. 40. V i t a m i n s Riboflavin (vitamin B2) overproducers include two yeast-like molds. 40. L-leucine. the control of the process involves the inhibition of phosphofructokinase by citric acid.4 billion. Higher pH values (e. Lvaline. Techniques similar to those described above for amino acid fermentations have yielded IMP titers of 27 g l21. 100. Citric acid is produced via the Embden-Meyerhof pathway and the first step of the tricarboxylic acid cycle. L-phenylalanine. The key to the fermentation is to avoid feedback repression by vitamin B12. These conditions prevent vitamin B12 synthesis and allow for the accumulation of the intermediate. or recombinant Bacillus subtilis strains that produce up to 30 g l21 riboflavin have been developed in recent years. denitrificans fermentation. Ltryptophan. converting cobinamide to the vitamin. L-tyrosine.

Industrial ethanol is mainly manufactured by fermentation. As a group that includes antibiotics. the fermentation can be continued to produce alcohol concentrations of 20% by volume. but these concentrations are attained only after months or years of fermentation. biopesticides and animal and plant growth factors. but some is produced from ethylene by the petrochemical industry. Alcohol Ethyl alcohol is a primary metabolite produced by fermentation of sugar. Such yeasts are able to convert n-paraffins to citric and isocitric acids in extremely high yields [150–170% (w/w) of substrate used]. The reactions are stereospecific. biomass) and crystalline cellulose directly to ethanol. microorganisms are extremely useful in carrying out biotransformation processes in which a 28 TIBTECH JANUARY 2000 (Vol. which would normally yield gluconic acid. the ultimate in specificity is exemplified by steroid bioconversions. Such a high concentration slows down growth and the fermentation ceases. all beverage alcohol is made by fermentation. Bacteria such as clostridia and Zymomonas are being re-examined for ethanol production after years of neglect.e. Fuel ethanol produced from biomass would provide relief from air pollution caused by the use of gasoline and would not contribute to the greenhouse effect. Under optimum conditions. can convert waste cellulose (i. coli has been converted into an excellent ethanol producer (43% yield. 18) . Other attributes include mild reaction conditions and the coupling of reactions. lactate. approximately 10– 12% ethanol by volume is obtained within five days. using a microorganism containing several enzymes working in s e r i e s . titers as high as 225 g l21 have been reached. Other clostridia produce acetate. with titers reaching about 100 g l21. they Bioconverting-organisms In addition to the multireaction sequences of fermentations. instead of citric acid. At present. an anaerobic thermophile. Alternative processes have been developed for the production of citric acid by Candida yeasts. acetone and butanol. Bioconversions are characterized by extremely high yields. E. With special yeasts. compound is converted into a structurally related product by one or a small number of enzymes contained in the cells5. Seconda ry metabol ites Microbially produced secondary metabolites6 are extremely important for health and nutrition. is the substrate. respectively. The low pH inactivates glucose oxidase. especially from hydrocarbons. Clostridium thermocellum. or a polysaccharide that can be depolymerized to a fermentable sugar. toxins. and will be used to produce these chemicals when the gobal petroleum supplies begin to become depleted. other medicinals. In approximately 4–5 days. whereas Kluyveromyces fragilis or Candida species can be used if lactose or a pentose. Saccharomyces cerevisiae is used for the fermentation of hexoses. approximately 90–100%. Bioconverting-organisms are known for practically every type of chemical reaction. v/v) by recombinant DNA techniques.oxalic and gluconic acids. the major proportion (80%) of the sugar is converted to citric acid.

and discover safer. there has been a virtual explosion of new and potent antibiotic molecules that have been of great use in medicine. such as monensin). Non-antibiotic agents In nature. The avermectins. viruses and parasites. Today. Antibiotics The best-known group of the secondary metabolites are the antibiotics7. microbial broths filled the void and microbial products increased in importance in the therapy of non-microbial diseases9. only a few promising structures were found. tetracycline. more potent and broaderspectrum compounds. (4) agents of symbiosis. lasalocid and salinomycin dominate the coccidiostat market and are also the chief growth promoters in use for ruminant animals. insects and plants. amoebae. transcription (rifamycin).ducing cultures (although.and 80-S ribosomes (puromycin and fusidic acid). but also for growth promo- 29 tion in farm animals and for the protection of plants. in order to: combat evolving pathogens. have high activity against helminths and arthropods. in nature. For years. agriculture and basic research. transcription by 80-S ribosomes (cyclohexamide). such as linear gramicidin.duced by certain restricted taxonomic groups of organ. cephalosporin and vancomycin) and cell membranes (surfactants including: polymyxin and amphotericin. Antibiotics are used not only for chemotherapy in human and veterinary medicine. cell wall synthesis (cycloserine. As new lead compounds became more and more difficult to find. translation by 70-S ribosomes (chloramphenicol. In the search for new antibiotics. they are essential for the survival of the producing organism). the antibiotic market was composed of 160 antibiotics and amounted to a world market value of ~US$23 billion. Despite the testing of thousands of synthetic compounds. fungi. improve pharmacological properties. Their targets include DNA replication (actinomycin. many of the new products are made chemically by modification of natural antibiotics via semisynthesis. In 1996. bleomycin and griseofulvin). microbially produced polyethers such as monensin. naturally resistant bacteria and fungi. lincomycin. channelforming ionophores. and previously susceptible microbes that have developed resistance.isms and are usually formed as mixtures of closely related members of a chemical family. erythromycin and streptomycin). are pro. functioning as: (1) sex hormones. coccidiostats and antihelminthics) came from the screening of synthesized compounds followed by molecular modification. transcription by 70. (3) competitive weapons against other bacteria. bacitracin. another group of streptomycete products with a market of more than US$1 billion per year. penicillin. The search for new antibiotics continues. and (5) effectors of differentiation. TIBTECH JANUARY 2000 (Vol. most pharmaceuticals that were used for non-infectious diseases were strictly synthetic products8. Secondary metabolites have no function in the growth of the pro. combat tumors. Similarly.g. and mobile carrier ionophores. Since 1940. (2) ionophores.have tremendous economic importance. secondary metabolites are important to the organisms that produce them. most therapeutics for nonmicrobial parasitic diseases in animals (e. 18) .

the two phases overlap. The precursor may also direct the fermentation towards the formation of one specific desirable product: this is known as directed biosynthesis. the timing depends on the nutritional environment presented to the culture. In such cases. spinosyns). rapamycin). In other fermentations. antihelminthics (avermectin).known enzyme inhibitors include: clavulanic acid. Stimulatory precursors include: methionine. a natural inhibitor of intestinal glucosidase.g. ruminant-growth promoters (monensin. uterocontractants (ergot alkaloids) and antitumor agents (doxorubicin. Resistance mechanisms that develop in producing microorganisms include enzymatic modification of the antibiotic. including lovastatin (also known as mevinolin) and pravastatin: fungal products that are used as cholesterol-lowering agents in humans and animals. daunorubicin. mitomycin. One huge success has been the statins. most secondary metabolite processes have a distinct growth phase (trophophase) followed by a production phase (idiophase). as an inducer in cephalosporin C formation. bioinsecticides (nikkomycin. plant-growth regulators (gibberellins). coccidiostats. precursors show no activity because their syntheses are not rate-limiting. Biopesticides Also in commercial or near-commercial use are biopesticides. A delay in antibiotic production until after trophophase helps the producing organism because the microbe is sometimes sensitive to its own antibiotic during growth. the liver and the intestinal wall of patients suffering from diabetes. however. FK-506. In its hydroxy acid form. and specific amino acids in the production of actinomycins and tyrocidins. Occasionally. immunosuppressants for organ transplants (cyclosporin A.logical activities were discovered by screening for inhibitors using simple enzymatic assays. lasalocid. Directed biosynthesis The manipulation of the culture media in any development program often involves the testing of hundreds of additives as possible limiting precursors of the desired product. alteration of the cellular target of the antibiotic and decreased uptake of the excreted antibiotic. valine. in tylosin production. and tryptophan for ergot-alkaloid production. a precursor that increases production of the secondary metabolite is found. a penicillinase-inhibitor that protects penicillin from inactivation by resistant pathogens. obesity and type IV hyperlipidemia. and acarbose. the growth rate. polyoxins). bioherbicides (bialaphos). including biofungicides (e.Many microbial products with important pharmaco.thesis in adipose tissue. salinomycin). In many fermentations. bleomycin). or both. Acarbose decreases hyperglycemia and triglyceride syn. which is produced by an actinomycete of the genus Actinoplanes. Tropophase and idiophase In batch culture. . anabolic agents in farm animals (zearelanone). kasugamycin. Other well. lovastatin is a potent competitive inhibitor of 3-hydroxy-3-methylglutaryl-coenzyme A reductase from liver. Examples of directed biosynthesis include the use of phenylacetic acid in the fermentation of benzylpenicillin.

B. the birth of recombinant DNA technology TIBTECH JANUARY 2000 (Vol. In 1972.ing decreases in costs have come about mainly by ran. Newer companies entered the scene in various niches such as biochemical engineering and downstream processing. mutations have had a major effect on the production of secondary metabolites. (2) elucidate the pathways of secondary metabolism. cerevisiae. Pichia pastoris. 18) propelled biotechnology to new heights and led to the establishment of a new industry. in vitro mutagenesis (protein engineering) and directed evolution of enzymes (applied molecular evolution). The use of recombinant microorganisms (Fig. described above. it is the chief factor responsible for the 100– 1000-fold increases obtained in the production of antibiotics from their initial discovery to the present time. cell immobilization (enzyme engineering). Antibiotic biosynthesis ends via the decay of antibiotic synthetases or because of feedback inhibition and repression of these enzymes. 1) provided the techniques and experience necessary for the successful application of higher organisms. subtilis. S. microbial physiology. such as mammalian and insect cell culture. coli11. These effects are usually due to the interaction of these compounds with the regulatory mechanisms existing in the fermentation organism.screening of additives has often revealed dramatic effects. bioreactor design. high-throughput screening for novel metabolites and strain improvement. 1980s and 1990s depended heavily upon the solid structure of industrial microbiology. Within four years of the discovery of recombinant DNA technology. Because the regulatory mechanisms are genetically determined. These tremendous increases in fermentation productivity and the result. This led to an explosion of investment activity in new companies. cell fusion. of which sales represent US$13. Mutation has also served to: (1) shift the proportion of metabolites produced in a fermentation broth to a more favorable distribution. and there are 29 . In addition to recombinant DNA technology. Hansenula polymorpha and Aspergillus niger. both stimulatory and inhibitory. downstream processing. The major microbial hosts for production of recombinant proteins are E. metabolic engineering. Modern microbial biotechnology Modern biotechnology is now over 25 years old10. Indeed. Recombinant microorganisms The revolutionary exploitation of microbial genetic discoveries in the 1970s. mainly dedicated to innovation via genetic approaches. modern microbial biotechnology encompasses fermentation.6 billion. bioreactor design and downstream processing. and transgenic animals and plants as hosts for the production of glycosylated recombinant proteins. Today. Progress The progress in biotechnology has been truly remarkable.dom mutagenesis and screening for higher-producing microbial strains. and (3) yield new compounds. of non-precursor molecules on the production of secondary metabolites. genetically engineered bacteria were making human insulin and human growth hormone.4 billion. biotechnology in the USA is represented by some 1300 companies with revenues of US$19.

erythropoietin. enzymes. which facilitates the transfer of an entire pathway in a single manipulation.1 billion. human growth hormone.7 billion. 1178 biotechnology companies existed with 45 000 employees. pathway genes are sometimes clustered. the first subunit vaccine on the market TIBTECH JANUARY 2000 (Vol. Even in fungi. recombinant DNA techniques are being used to introduce genes for the synthesis of one product into producers of other antibiotics or into non-producing strains (combinatorial biosynthesis). antihemophelia factor. European biotechnology moved rapidly in the 1990s. but also came down with the disease. The number of biotechnology companies in Canada reached 282 in 1998. after years of lagging behind and. food and beverage compa. (Electron micrograph taken by Erika Hartweig. bioconversion and secondary metabolites.nies. and revenues of US$3.FEATURES FEATURES was that of hepatitis B virus surface antigen produced in yeast. Japan’s biotechnology sales were approximately US$10 billion. such as hormones. V a c c i n e s Vaccine production is another important part of the new technology. a-. bar = 1 mm. Through reversion of the attenuated pathogen. cally engineered products that have been approved for use in the USA are human insulin. The major thrust of recombinant DNA technology has been in the area of rare mammalian peptides.and g-interferons. recombinant DNA techniques have made a significant impact on the production of vitamins. and bovine somatotropin. 18) . interleukin-2. Figure 1 Escherichia coli: the workhorse of modern microbial biotechnology.) approximately 153 000 employees. mainly by established pharmaceutical. nucleotides. such as the penicillin genes in Penicillium or the aflatoxin genes in Aspergillus. amino acids. epidermal growth factor and other growth factors. granulocyte-colony stimulating factor. some individuals receiving the conventional vaccine not only failed to be protected.ciated with conventional vaccines. Among those geneti30 Combinatorial biosynthesis As mentioned previously. For the discovery of new or modified secondary products. in 1998. employing 10 000 workers and with revenues of approximately US$1. granulocytemacrophage-colony stimulating factor. growth factors. b. antibodies and biological response modifiers12. Most microbial biosynthetic pathways are encoded by clustered genes. The great contribution made by recombinant vaccines is the elimination of the tragic problems asso.

thuringiensis. metabolic diseases and cancer. lipases. B. approximately 300 times more active TIBTECH JANUARY 2000 (Vol. gastrointestinal and rheumatic disorders. However. Recombinant therapeutic enzymes already have a market value of over US$2 billion. thuringiensis preparations are highly potent. recombinant chymosin for cheese manufacture and recombinant lipase for use in detergents. 18) . The activity of the insecticidal bacterium. Crystals and spores have been applied to plants for many years to protect them against lepidopteran insects. is caused by its crystal protein produced during sporulation. B. carbohydrases. Two bacteria have had a major 31 FEATURES influence: Agrobacterium tumefaciens.FEATURES Enzyme production The production of enzymes by fermentation was an established business before modern microbial biotechnology. for example. Agriculture Industrial microbiology through genetic engineering and its associated disciplines has brought about a revolution in agriculture. along with genes directing the plant to form opines (nutritional factors required by the bacterium that it cannot produce by itself ). the development of a particleacceleration gun. a problem bypassed by. Important enzymes are proteases. tumefaciens are present on its tumor-inducing (Ti) plasmid. human DNAase and Cerozyme.6 billion. However. recombinant DNA methodology was so perfectly suited to the improvement of enzymeproduction technology that it was almost immediately used by companies involved in manufacturing enzymes. being used for thromboses. They include tissue plasminogen activator. a bacterium that normally produces crown gall tumors on dicotyledonous plants. The Ti vector has been exceedingly valuable for introducing foreign genes into dicotyledonous plants for production of transgenic plants. Industrial enzymes have now reached an annual market of US$1. The tumor-forming genes of A. the Ti plasmid is not very successful for transferring genes into monocotyledonous plants. which shoots DNA-coated metal particles into plant cells. and Bacillus thuringiensis. an insecticidal bacterium.

(1988) Contributions of genetics to the production and discovery of microbial pharmaceuticals.boulnois@astrazeneca.oped bioinsecticides include insect viruses.g. Marcel Dekker References 1 Demain. 16. eds). pp. Pure Appl. eds). and will continue to make.L. M Biotechnology has already played a major role in modern medicine discovery. Biotechnol. and the past few decades have seen enormous advances in our ability to treat. F. M. H. technology. are also available. 16. drive and dedication to solve the problems of evolving diseases (e.R. by converting urban. S. ed. chemical pesticides against plant viruses were never available. (1997) Biotransformations. In Recombinant Microbes for Industrial and Agricul. In the modern biotechnology era. and gene ther.com) is at AstraZeneca Pharmaceuticals. Successful therapies range from the treatment of acute infections with powerful antibiotics and the availability of anaesthetics for surgery and management. (1992) Microbial secondary metabolism: a new theoretical frontier for academia. resistant to herbicides. major support by governments and international agencies.). manage and prevent a large number of diseases.apy is just round the corner. antibioticresistance development and environmental pollution. Recently devel. major challenges remain. 3–4 10 Cohen. as well as an understanding and supportive public. 15. A. pp. UK SK10 4TG.J. ed. Nat. American Society of Microbiology (ASM) Press 12 Demain. We must use our brains.J. pp. a new opportunity for industry. Macclesfield. Biotechnol. industrial and agricultural wastes into resources such as liquid fuel.. 173–204 11 Swartz. (1979) The transplantation and manipulation of genes in microorganisms. 243–252 5 Kieslich. In A Century of Mycology (Sutton. A. odern medicines have improved the life of humankind. Biotechnol. (1994) Contributions of recombinant microbes and their potential.L.J. Rev. and Imanaka.L. 27–46. These efforts will require continued interaction between different disciplines.C. T. Chem.uloviruses. Conclusion Although most of the early promises of biotechnology have been achieved. delivering a range of new treatments and vaccines in its own right. (1996) Escherichia coli recombinant DNA technology. a major contribution to the health of mankind by providing new insights into disease and acting as a pivotal enabler for the drug-discovery process. pp. In Escherichia coli and Salmonella: Cellular and Molecular Biology 2nd edn (Neidhardt.pressing viral-coat-protein genes in plants.. such as bac. Marcel Dekker 8 Demain. Crit. Rev.. 233– 254. thuringiensis toxin gene in the plant. 291–301 Drug discovery in the new millennium: the pivotal role of biotechnology Graham J. 3–23.). A. that are engineered to produce arthropod toxins.N. (1997) Biotechnology of Antibiotics. through risk reduction in cardiovascular disease to the prevention of a range of infectious diseases via vaccination. The Harvey Lectures 74.R. . In Secondary Metabolites: Their Function and Evolution (Chadwick. A. Adv. 60. as are virus-resistant plants produced by ex. 833–836 3 Jetten. Cambridge University Press 9 Demain.on a molar basis than synthetic pyrethroids and 80 000 times more active than organophosphate insecticides..tural Applications (Murooka.L. J. A. 1693–1711. 2 Demain. plants resistant to insects have been produced by expressing forms of the B. AIDS). In Fungal Biotechnology (Anke. pp. Despite these impressive G. A.L.S. (1990) Achievements in microbial technology. T. 2nd edn. ed.. 73–103 4 Sahm.). T. A. (1995) Recent advances in the physiology and genetics of amino acid-producing bacteria. (1995) Metabolic design in amino acid producing bacterium Corynebacterium glutamicum. Transgenic plants. established diseases (cancer and parasitic infection). W. The available techniques are diverse and changing rapidly: selecting and integrating the best approach is the key to success. (1998) Microbial natural products: alive and well in 1998. 8. FEMS Microbiol. (1996) Fungal secondary metabolism: regulation and functions. Boulnois Biotechnology has made. John Wiley & Sons 7 Strohl. and Sinskey. K. et al. Chapman & Hall 6 Demain. Interestingly. 297–399. J.L. and Whelan. D. Alderley Park. et al. B. Boulnois (graham.

has changed to a molecular target approach in which in vitro screening of compounds against purified.advances.eral but particularly by molecular biology. This change has come about as a consequence of better and ever-improving knowledge of the molecular basis .cines are discovered. as an early part of the discovery process. recombinant proteins or genetically modified cell lines is carried out with a high throughput. there are many diseases for which treatments are lacking or are imperfect. In short. We have also seen a change in the way new medi. The reliance on testing new synthetic organic molecules in animals or in whole-organ preparations. the opportunities for new treatments is huge. and we are a long way from eradicating or even reducing the prevalence of many debilitating conditions. driven by biotechnology in gen.

TIBTECH JANUARY 2000 (Vol. All rights reserved. 18) 0167-7799/00/$ – see front matter © 2000 Elsevier Science Ltd. PII: S0167-7799(99)01393-1 31 .